Peptide-crosslinked Protein-imprinted Polymers And Preparation Method And Applications Thereof

Patent metadata

Field	Value
Publication number	US-2021388123-A1
Application number	US-201916954644-A
Country	US
Kind code	A1
Filing date	Apr 19, 2019
Priority date	Feb 28, 2019
Publication date	Dec 16, 2021
Grant date	—

Abstract

A peptide-crosslinked protein-imprinted polymer, preparation method, and application thereof. One method comprises: 1) dissolving a main monomer, functional monomers, a peptide crosslinking agent, and a template protein in an aqueous solution to obtain a mixed solution; 2) adding an initiator or initiator system to the mixed solution to initiate the polymerization when the peptide crosslinking agent exists in a helix conformation to obtain a polymer; 3) eluting the template protein when the peptide chain exists in a coil conformation to obtain a peptide-crosslinked protein-imprinted polymer. The peptide crosslinking agent is a peptide with a polymerizable double bond at its both ends, and being capable of undergoing helix-coil transition. The polypeptide crosslinking agent is a polypeptide having an amino acid sequence which has a polymerizable double bond at its both ends, being capable of undergoing a helix-coil conformational transformation. The polypeptide cross-linked protein molecule-imprinted polymer disclosed in the invention not only can completely remove the template protein under mild conditions, but also can significantly improve the imprint effect of the protein molecule-imprinted polymer.

First claim

What is claimed is:

1 . A method of preparing a peptide-crosslinked protein-imprinted polymer comprising:
dissolving a main monomer, functional monomers, a peptide crosslinking agent, and a template protein in an aqueous solution to obtain a mixed solution; adding an initiator or initiator system to the mixed solution to initiate the polymerization while the peptide crosslinking agent exists in a helix conformation, thereby obtaining a polymer; eluting the template protein while the peptide chain exists in a coil conformation, thereby obtaining a peptide-crosslinked protein-imprinted polymer; wherein the peptide crosslinking agent is a peptide with a polymerizable double bond at both ends, the peptide crosslinking agent being capable of undergoing a helix-coil transition.

2 . The method according to claim 1 , wherein the peptide crosslinking agent comprises a quantity of amino acids from 2 to 100.

3 . The method according to claim 1 , wherein the peptide crosslinking agent comprises a quantity of amino acids from 5 to 30.

4 . The method according to claim 3 , wherein the aqueous solution comprises one of a phosphate buffer, a Tris buffer, and a NaClO 4 solution.

5 . The method according to claim 1 , wherein the aqueous solution comprises one of a phosphate buffer, a Tris buffer, and a NaClO 4 solution.

6 . The method according to claim 1 , wherein the mixed solution comprises a mass ratio of ranges of the main monomer:the functional monomers:the peptide crosslinking agent:the template protein of (10-30):(0.5-2):(5-20):(5-20).

7 . The method according to claim 1 , wherein the initiator is selected from the group consisting of: a combination of ammonium persulfate solution and tetramethylethylenediamine, a combination of ammonium persulfate solution and sodium bisulfite, a combination of potassium persulfate solution and tetramethylethylenediamine, and a combination of potassium persulfate solution and sodium bisulfite.

8 . The method according to claim 1 , further comprising detecting a conformational state of the peptide crosslinking agent by circular dichroism (CD), in which:
a CD spectrum presents a negative peak at 222 nm and 208 nm, and a positive peak at 190 nm±5 nm representative of a helix conformation; and a CD spectrum presents a negative peak at 199 nm, and a positive peak at 220 nm±5 nm representative of a coil conformation.

9 . The peptide-crosslinked protein-imprinted polymer prepared by the method according to claim 1 , wherein the peptide-crosslinked protein-imprinted polymer comprises a polypeptide-crosslinked protein-imprinted polymer.

10 . An application of the polypeptide-crosslinked protein-imprinted polymer according to claim 9 in protein enrichment, separation, or purification.

11 . The application according to claim 10 , wherein the polypeptide-crosslinked protein-imprinted polymer is used to:
adsorb target protein while the peptide chain exists in a helix conformation and elute target protein when the peptide chain exists in a coil conformation.

Assignees

• Univ Nankai

Inventors

• Zhang Yongjun
• Xu Rong
• Guan Ying

Classifications

• C08F289/00

  Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds not provided for in groups C08F251/00 - C08F287/00 · CPC title

• C08F220/06

  Acrylic acid; Methacrylic acid; Metal salts or ammonium salts thereof · CPC title

• C07K17/08

  the carrier being a synthetic polymer · CPC title

• C08F290/065

  Polyamides; Polyesteramides; Polyimides · CPC title

• C08F220/54

  Amides {, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide} · CPC title